TW201444824A - Quinazolines as kinase inhibitors - Google Patents

Abstract

Disclosed are compounds that are inhibitors of RIP2 kinase and methods of making and using the same.

Description

Quinazolines as kinase inhibitors

The present invention relates to quinazolines for inhibiting RIP2 kinase and methods of making and using same.

Receptor-interacting protein-2 (RIP2) kinase, also known as CARD3, RICK, CARDIAK or RIPK2, is a TKL family of serine/tyrosine protein kinases involved in innate immune signaling. The RIP2 kinase system consists of an N-terminal kinase domain and a C-terminal caspase-recruitment domain (CARD) linked via an intermediate (IM) region. ((1998) J. Biol. Chem. 273, 12296-12300; (1998) Current Biology 8, 885-889; and (1998) J. Biol. Chem. 273, 16968-16975). The CARD region of RIP2 kinase mediates interactions with other CARD-containing proteins, such as NOD1 and NOD2 ((2000) J. Biol. Chem. 275, 27823-27831 and (2001) EMBO reports 2, 736-742). NOD1 and NOD2 are a cytoplasmic receptor that plays a key role in innate immune surveillance. It identifies Gram-positive and Gram-negative bacterial pathogens and is activated by specific peptidoglycan motifs, diaminopimelic acid (also known as DAP) and cell wall quinone dipeptide (MDP) (2007 J Immunol 178, 2380-2386).

Upon activation, RIP2 kinase binds to NOD1 or NOD2 and appears to function primarily as a molecular framework for the aggregation of other kinases involved in the activation of NF-κB and mitogen-activated protein kinases (TAK1, IKKα/β/γ) ( 2006) Nature Reviews Immunology 6, 9-20). RIP2 kinase performs K63-linked polyubiquitination on lysine-209, which promotes TAK1 Recruitment ((2008) EMBO Journal 27, 373-383). This post-translational modification is necessary for signal delivery because the variation of this residue prevents NF-kB activation of the NOD1/2 vector. RIP2 kinase also autophosphorylates on serine-176, and possibly other residues ((2006) Cellular Signalling 18, 2223-2229). Studies using kinase inactivating mutants (K47A) and non-selective small molecule inhibitors have demonstrated that RIP2 kinase activity is important for regulating RIP2 kinase expression and signal transmission stability ((2007) Biochem J 404, 179-190 and (2009) J. Biol. Chem. 284, 19183-19188).

RIP2-dependent signal transmission regulation abnormalities are associated with autologous inflammatory diseases. Functional acquired mutations in the NACHT-region of NOD2 result in Blau syndrome characterized by uveitis, skin inflammation and arthritis, early onset sarcoidosis, pediatric granulomatous disease (2001) Nature Genetics 29, 19-20; (2005 Journal of Rheumatology 32, 373-375; (2005) Current Rheumatology Reports 7, 427-433; (2005) Blood 105, 1195-1197; (2005) European Journal of Human Genetics 13, 742-747; (2006) American Journal of Ophthalmology 142, 1089 -1092; (2006) Arthritis & Rheumatism 54, 3337-3344; (2009) Arthritis & Rheumatism 60, 1797-1803; and (2010) Rheumatology 49, 194-196). Mutations in the LRR-region of NOD2 are closely related to susceptibility to Crohn's disease ((2002) Am. J. Hum. Genet. 70, 845-857; (2004) European Journal of Human Genetics 12, 206-212; (2008) Mucosal Immunology (2008) 1 (Suppl 1), S5-S9.1, S5-S9; (2008) Inflammatory Bowel Diseases 14, 295-302; (2008) Experimental Dermatology 17, 1057-1058; (2008) British Medical Bulletin 87, 17-30; (2009) Inflammatory Bowel Diseases 15, 1145-1154 and (2009) Microbes and Infection 11, 912-918). The mutation of NOD1 is associated with asthma ((2005) Hum. Mol. Genet. 14, 935-941) and early onset and extraintestinal inflammatory bowel disease ((2005) Hum. Mol. Genet. 14, 1245-1250). Genetic and functional studies have also shown that RIP2-dependent signaling is involved in a variety of other granulomatous disorders, such as sarcoidosis (2009) Journal of Clinical Immunology 29, 78-89 and (2006) Sarcoidosis Vasculitis and Diffuse Lung Diseases 23, 23-29) and Wegener's granulomatosis ((2009) Diagnostic Pathology 4, 23).

A potent, selective small molecule inhibitor of RIP2 kinase activity should block RIP2-dependent pre-inflammatory signaling and thus provide auto-inflammatory and/or autoimmune diseases characterized by increased RIP2 kinase activity and/or abnormal regulation. Treatment benefits.

The present invention relates to a compound selected from the group consisting of 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7- 2-methoxyethoxy)quinazolin-4-amine: 6-(T-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazolin-4-amine of the formula: 6-(T-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-propoxyquinazolin-4-amine of the formula: And 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-((tetrahydrofuran-2-yl)methoxy having the following formula Quinazoline-4-amine: Or a salt thereof, especially a pharmaceutically acceptable salt thereof.

Accordingly, the present invention relates to a method of inhibiting RIP2 kinase, which comprises contacting a cell with a compound of the invention or a salt thereof, particularly a pharmaceutically acceptable salt.

The invention further relates to a method of treating a RIP2 kinase-mediated disease or condition comprising administering a therapeutically effective amount of a compound of the invention, or a salt thereof, particularly a pharmaceutically acceptable salt thereof, to a patient in need thereof (human, or other mammals, especially humans). The invention further relates to the use of a compound of the invention or a pharmaceutical composition comprising a compound of the invention for inhibiting a RIP2 kinase and/or RIP2 kinase-mediated disease or condition.

Examples of diseases or conditions of RIP2 kinase-mediated include uveitis, Crohn's disease, ulcerative colitis, early-onset extraintestinal inflammatory bowel disease, and granulomatous diseases such as sarcoidosis, Blau syndrome, early onset sarcoidosis And Wegener's granulomatosis.

The invention further relates to a pharmaceutical composition comprising a compound of the invention or a salt thereof, in particular a pharmaceutically acceptable salt, and a pharmaceutically acceptable excipient. In particular, the invention relates to a pharmaceutical composition for the treatment of a RIP2 kinase-mediated disease or condition, wherein the composition comprises a compound of the invention or a salt thereof, in particular a pharmaceutically acceptable salt thereof, and one or more pharmaceuticals Excipients are acceptable.

Detailed description of the invention

As used herein, the term "compound of the invention" refers to any of the compounds as defined herein, ie, any salt or non-salt form (eg, in the form of a free acid or base, or a salt, especially pharmaceutically Accept salt) and any physical form thereof (for example, including non-solid forms) (eg, in liquid or semi-solid form) and in solid form (eg, amorphous or crystalline form, specific polymorph form, solvate form, including hydrated forms (eg, mono-, di-, and hemi-hydrate), and various Mixtures of Forms (Hydrates of Salts). In particular, it is to be understood that the present invention encompasses the compounds of the invention which are free bases and salts thereof, for example, pharmaceutically acceptable salts thereof. In one embodiment, the invention relates to The compound of the invention is in the form of a free base. In a further embodiment, the invention relates to a compound of the invention in the form of a salt, especially a pharmaceutically acceptable salt.

Those skilled in the art will also appreciate that the pyrazolyl group present in the compounds of the present invention may exist as tautomeric pyrazolyl isomers represented by formula (IA) and formula (IB).

It will be appreciated that the pyrazolyl group produced may be designated as a 3,4-dimethyl-1H-pyrazol-5-yl group or a 4,5-dimethyl-1H-pyrazol-3-yl group. group. It will be understood that any reference to the named compounds of the invention is intended to encompass all tautomers of the named compound and any mixtures of tautomers of the named compound. For example, the compound name 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazolin-4-amine The compound covers the compound 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazolin-4-amine and 6 -(T-butylsulfonyl)-N-(3,4-dimethyl-1H-pyrazol-5-yl)-7-ethoxyquinazolin-4-amine, and mixtures thereof. All tautomeric forms of the compounds described herein are intended to be encompassed within the scope of the invention.

The compounds of the invention may contain one or more asymmetric centers (also known as palm centers) and may therefore be present in the form of individual mirror image isomers, diastereomers or other stereoisomers or mixtures thereof. The center of the palm, such as palmitic carbon, may also be present in the compounds of the invention. Compounds, compound names or structures are intended to encompass all individual stereotypes when stereochemistry of the palm center in the presence of a compound of the invention or any chemical structure illustrated herein is not indicated. Isomers and all mixtures thereof. Thus, a compound of the invention containing one or more centers of palms may be present as a racemic mixture, a mixture rich in mirror phase isomers or a mirror-isomeric pure individual stereoisomer.

Individual stereoisomers of a compound of the invention containing one or more asymmetric centers can be resolved by methods known to those skilled in the art. For example, the resolution can be (1) by formation of diastereomeric salts, complexes or other derivatives; (2) by selective reaction with stereoisomer-specific reagents, by enzyme properties Oxidizing or reducing; or (3) by gas phase-liquid phase or liquid phase chromatography in a palm environment, such as on a palmitic carrier, such as with a binding pair of palmitic ligands or in the palm In the presence of a solvent, it is carried out. Those skilled in the art will appreciate that when the desired stereoisomer is converted to another chemical entity by one of the separation processes described above, an additional step is required to dissociate the desired form. Alternatively, specific stereoisomers can be converted to another mirror image by asymmetric synthesis using optically active reagents, matrices, catalysts or solvents, or by asymmetric transformation. And synthesize.

It will be appreciated that the solid form of the compounds of the invention may exist in crystalline form, in amorphous form or as a mixture thereof. These crystal forms may also have pleomorphism (i.e., the ability to produce different crystal forms). These crystal forms are typically referred to as "polymorphs." Polymorphs have the same chemical composition but differ in the nature of the deposition, geometric alignment, and other crystalline solid states. Polymorphs can therefore have different physical properties such as shape, density, hardness, deformability, stability and solubility properties. Polymorphs typically have different melting points; IR spectra and X-ray powder diffraction modes, which can be used for identification. It will be understood by those of ordinary skill in the art that different polymorphs can be produced, for example, by varying and adjusting the conditions used to crystallize/crystallize the compound.

It is well known and understood by those skilled in the art that the apparatus used to obtain the X-ray powder diffraction (PXRD) mode, humidity, temperature, orientation of powder crystals, and other parameters may be in a linear position in appearance, density, and diffraction mode. Cause some variability. It is "substantially identical" to the X-ray powder diffraction mode of the figure provided in the text. A person familiar with the art should be regarded as a PXRD mode representing a compound having the same crystal form as the compound of the PXRD mode proposed in the figure. formula. For example, the PXRD mode may be the same as FIG. 1, or more likely it may be slightly different. This PXRD mode may not require lines showing the diffraction patterns shown in the text, and/or may exhibit some variation in the appearance, density, or positional movement of the lines due to differences in the conditions involved in obtaining the data. Those skilled in the art, by comparing their PXRD patterns, can determine whether a sample of the crystalline compound has the same form, or a different form, as described herein. For example, those skilled in the art can use 2,6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquin. The PXRD pattern of the crystalline form of the oxazolin-4-amine (free base) overlaps with the PXRD pattern of Figure 1, and using the expertise and knowledge of this technique, it is easy to determine whether the PXRD mode of this sample is substantially the same as in Figure 1. The PXRD mode is consistent. If the PXRD mode is substantially identical to that of Figure 1, the sample can be readily and correctly identified as having 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H as described herein. -Pyrazol-3-yl)-7-ethoxyquinazolin-4-amine (free base) has the same crystalline form. Similarly, those skilled in the art will be able to determine whether the particular diffraction angle (indicated by 2θ) obtained by the PXRD mode is at approximately the same position of the value.

Because of their potential medical use, the salts of the compounds of this invention are pharmaceutically acceptable salts. Suitable pharmaceutically acceptable salts include acid or base addition salts such as those by Berge, Bighley and Monkhouse J. Pharm. Sci (1977) 66, pp 1-19 and "Pharmaceutical Salts: Properties, Selection, and Use". , Second Edition, "PHStahl and CGWermuth (eds.), Wiley, Hoboken, NJ, US (2011).

The term "pharmaceutically acceptable" means the compounds, substances, compositions and dosage forms which, within the scope of good medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation or other problems. Or a complication with a reasonable interest/risk ratio.

"Pharmaceutically acceptable salt" means a compound suitable for medical use. Salts and solvate forms (e.g., hydrates and hydrates of the hydrates) of the compounds of the invention suitable for use in pharmaceuticals are those in which the relative ionic or combined solvents are pharmaceutically acceptable. However, salts and solvates having non-pharmaceutically acceptable relative ionic or binding solvents are within the scope of the invention, for example, as intermediates in the preparation of other compounds of the invention and their salts and solvates.

When the compound of the invention is a base (containing a basic group), the desired salt form can be prepared by any suitable method known in the art, including treatment of the free base with an acid. Examples of pharmaceutically acceptable acid addition salts include acetate, adipate, ascorbate, aspartate, besylate, benzoate, camphorate, camphorsulfonate, citric acid Salt, hexanoate, octoate, carbonate, bicarbonate, cinnamate, citrate, cyclohexylamine sulfonate, lauryl sulfate (etolate), B-1,2-di Sulfonate (ethanedisulfonate), ethanesulfonate, formate, fumarate, galactose salt (mucosate), gentisate (2,5-dihydroxybenzoate) , glucoheptonate, gluconate, glucuronate, glutamed acid salt, glutarate, glycerol phosphate, glycolate, hippurate, hydrochloride, hydrobromide, Hydroiodide, isobutyrate, lactate, lacturonate, laurate, maleate, malate, malonate, mandelate, methanesulfonate, naphthalene-1,5 -disulfonate, naphthalenesulfonate, nicotinic acid salt, nitrate, oleate, oxalate, palmitate, pamoate, phosphate, diphosphate, propionate, coke bromide Amine, salicylate, azelaic acid Salt, stearate, succinate, sulfate, tartrate, thiocyanate, tosylate, undecylenate, 1-hydroxy-2-naphthate, 2,2-dichloroacetic acid Salt, 2-hydroxyethanesulfonate, 2-ketoglutarate, 4-acetamide benzoate and 4-aminosalicylate. Non-pharmaceutically acceptable salts, such as trifluoroacetic acid, can be used in the isolation of the compounds of the invention and are included within the scope of the invention.

When the compound of the invention is an acid (containing an acidic group), the desired salt form can be prepared by any suitable method known in the art, including treatment of the free acid with a base. Examples of pharmaceutically acceptable base addition salts include ammonium salts, lithium salts, sodium salts, potassium salts, calcium salts, magnesium salts, aluminum salts, zinc salts, trimethylamine, triethylamine, morphine, pyridine, and piperazine. Pyridine, picoline, dicyclohexylamine, N,N'-dibenzylethylenediamine, 2-hydroxyethylamine, bis-(2-hydroxyethyl)amine, tris-(2-hydroxyethyl) Amine, procaine, diphenylmethylpiperidine, dehydrofungsamine, glucosamine, N-methylglucamine, collidine, quinine, quinoline, lysine and essence Amino acid. In one embodiment, the pharmaceutically acceptable base addition salt is a sodium salt.

Particular compounds of the invention may form a salt with one or more equivalents of an acid if the compound contains a basic group. All possible stoichiometric and non-stoichiometric salt forms are included within the scope of the invention.

The invention also provides a pharmaceutically acceptable salt which converts a pharmaceutically acceptable salt of a compound of the invention to another compound of the invention.

If the basic compound is isolated as a salt, the corresponding free acid or free base form of the compound can be prepared by any suitable method known in the art.

For solvates of the compounds of the invention, including solvates of the salts of the compounds of the invention in crystalline form, it will be understood by those skilled in the art that pharmaceutically acceptable solvates can be formed in which the solvent molecules are incorporated into the crystal lattice during crystallization. . Solvates may include non-aqueous solvents such as ethanol, isopropanol, DMSO, acetic acid, ethanolamine, and EtOAc, or they may include water as a solvent, incorporated into the crystal lattice. Water in which the solvent is incorporated into the crystal lattice is typically referred to as a "hydrate." Hydrates include stoichiometric hydrates as well as compositions containing varying amounts of water. The invention includes all such solvates, especially hydrates.

Since the compounds of the invention are intended for use in pharmaceutical compositions, it will be readily appreciated that they are preferably each provided in substantially pure form, such as at least 60% pure, more suitably at least 75% pure and preferably At least 85%, especially at least 98% purity (% by weight based on weight). Preparations of impure compounds can be used to prepare the more pure forms used in pharmaceutical compositions.

General synthetic method

The compounds of the present invention can be prepared by using the synthetic procedures illustrated in the schemes below or by utilizing the knowledge of an organic chemist skilled in the art. The syntheses provided in these schemes, with suitable precursors, can be applied to the manufacture of compounds of the invention having various substituent groups which are suitably (if desired) protected to achieve the reaction phase outlined herein. Capacitance. Subsequent deprotection (when needed) yields compounds of the general disclosed nature. When the scheme is shown only as a compound of the formula, it is an illustration of a process that can be used to make the compounds of the invention.

The substitution of C6 can be carried out before the pyrazolyl group is placed. Palladium catalyzed coupling of mercaptan with 6-iodoquinazolinone gives a sulfide which can subsequently be oxidized to ruthenium. Chlorination with POCl ₃ or SOCl ₂ gives 4-chloroquinazoline.

The aniline/amine can be reacted with 4-chloro-quinazoline under basic or acidic conditions to give the 4-aminoquinazoline which can be the final compound or used as an intermediate for further synthesis.

Preparation of certain compounds of the invention, alternatively, may be prepared by heating from 6-bromo-7-fluoroquinazolin-4-ol via the reaction with a suitable alcohol in the presence of a base. 6-bromo-7-alkoxyquinazolin-4-ol. Subsequent chlorination and displacement with amine/aniline gives 4-amino-6-bromo-7-alkoxyquinazoline. These compounds are further reacted with a thiol or thiolate in the presence of a suitable palladium catalyst, ligand and base to give 4-amino-6-alkylthio-7-alkoxyquinazoline. . Oxidation will give the 4-amino-6-sulfonyl-7 alkoxyquinazoline, which may be the final compound or used as an intermediate for further chemical reactions.

Process 3

The preparation of certain of the compounds of the invention can be carried out from 7-fluoro-6-sulfonyl-4-quinazolinone. The synthesis of these intermediates begins with bromination of 4-fluoro-2-aminobenzoic acid followed by in situ condensation with formamidine acetate. Palladium catalyzed coupling with a thiol to give a sulfide which is subsequently oxidized to ruthenium.

The substitution of the fluorine substituent of the alkoxy group can be carried out by treatment with a suitable alcohol and sodium butoxide.

A particular compound of the invention is 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazoline-4- Amine (as the free base). In a further embodiment, the particular compound of the invention is 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxy A quinazolin-4-amine or a salt thereof. In a further embodiment, the particular compound of the invention is 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxy A quinazolin-4-amine or a pharmaceutically acceptable salt thereof. In a further embodiment, the particular compound of the invention is 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7 in crystalline form -ethoxy quinazolin-4-amine, characterized It is the PXRD mode of Figure 1. In yet another embodiment, the particular compound of the invention is 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)- in crystalline form. 7-Ethoxyquinazolin-4-amine characterized by the diffraction data of Table 1.

The compounds of the invention are inhibitors of RIP2 kinase. Thus, in one embodiment, the invention relates to a method of inhibiting RIP2 kinase comprising contacting a cell with a compound of the invention. In a further embodiment, the invention relates to a method of treating a RIP2 kinase-mediated disease or condition comprising administering a therapeutically effective amount of a compound of the invention to a human in need thereof.

In a further particular embodiment, the invention relates to a method of treating a RIP2 kinase-mediated disease or condition comprising administering a therapeutically effective amount of 6-(t-butylsulfonyl)-N-(4, 5-Dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazolin-4-amine or a salt thereof is administered to a human in need thereof.

The compounds of the invention are particularly useful for the treatment of a RIP2 kinase-mediated disease or condition, particularly where a disease or condition that inhibits the availability of RIP2 kinase. Examples of such RIP2 kinase-mediated diseases or conditions include uveitis, interleukin-1 converting enzyme (ICE, also known as apoptotic protease-1), fever syndrome (ICE fever), dermatitis, acute lung Injury, type 2 diabetes, arthritis (especially rheumatoid arthritis), inflammatory bowel disease (such as ulcerative colitis and Crohn's disease), early onset inflammatory bowel disease, extraintestinal inflammatory bowel disease, In response to ischemic reperfusion (especially kidney), liver disease (nonalcoholic steatohepatitis, alcoholic steatohepatitis and autologous) in solid organs of ischemia caused by cardiac surgery, organ transplantation, sepsis and other injuries Immune hepatitis), allergic diseases (such as asthma), transplant reactions (such as graft versus host disease), autoimmune diseases (such as systemic lupus erythematosus and multiple sclerosis), and granulomatous disorders (such as sarcoidosis, Blau syndrome) , early onset sarcoidosis, Wegener's granulomatosis and interstitial lung disease).

The compounds of the invention are particularly useful for the treatment of uveitis, ICE fever, Blau syndrome, early onset sarcoidosis, ulcerative colitis, Crohn's disease, Wegener's granulomatosis and sarcoidosis. A disease or condition that treats RIP2 kinase-mediated, or more broadly, diseases that treat the immune vector include, but are not limited to, allergic diseases, autoimmune diseases, prevention of transplant rejection, and the like, Using the compounds of the invention as a single treatment, or in a double or multiple combination, in particular for the treatment of refractory cases, for example in combination with other anti-inflammatory and/or anti-TNF agents, which may be known as locked in the art Administration is carried out in a therapeutically effective amount.

The compounds of the invention may be used alone or in combination with other therapeutic agents. Combination therapies according to the invention thus include administration of at least one compound of the invention, and the use of at least one other therapeutically active agent. Preferably, the combination therapy according to the invention comprises administering at least one compound of the invention, and at least one other therapeutically active agent. The compounds of the invention and the additional therapeutically active agents may be administered together or separately as a single pharmaceutical composition, and when administered separately, may be administered simultaneously or sequentially in any order. The amount of the compound of the invention and the additional therapeutically active agent, as well as the relative time of administration, will be selected to achieve the desired combination therapeutic effect. Thus, in another aspect, a composition comprising a compound of the invention together with one or more additional therapeutically active agents is provided. In another aspect, the invention provides 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazoline- A composition of 4-amine or a pharmaceutically acceptable salt thereof, together with one or more other therapeutically active agents.

Thus in one aspect, the compounds of the invention and pharmaceutical compositions comprising the compounds of the invention are useful in combination with one or more other therapeutically active agents, such as anti-inflammatory agents and/or anti-TNF agents.

The compounds of the invention may be combined with corticosteroids and/or anti-TNF agents for the treatment of Blau syndrome, early onset sarcoidosis; or in combination with anti-TNF biologics for the treatment of Crohn's disease; or with 5-ASA (Mesalazine) (mesalamine) or sulfasalazine in combination for the treatment of ulcerative colitis; or in combination with low-dose corticosteroids and/or methotrexate for the treatment of Wegener's granulomatosis or sarcoidosis or Pulmonary lung disease; or in combination with biological agents (such as anti-TNF, anti-IL-6, etc.) for the treatment of rheumatoid arthritis; or in combination with anti-IL6 and / or methotrexate for the treatment of ICE fever .

Examples of anti-inflammatory agents include 5-aminosalicylic acid and mesalazine preparations, sulfasalazine, hydroxycloroquine, thiopurine (azathioprine) (azathioprin), mercaptopurin, methotrexate, cyclophosphamide, cyclosporine, JAK inhibitor (tofacitinib), corticosteroids, especially low dose corticosteroids (eg, prednisone and bundesonide and anti-inflammatory biologics such as anti-IL6R mAb (Actemra® (tocilizumab)), anti-IL6 biologics, anti-IL1 Or IL12 or IL23 biologics (ustekinumab (Stelara®)), anti-integrin agents (natalizumab (Tysabri®)), anti-CD20 mAbs (rituximab) Other agents such as rituximab (Rituxan®) and ofatumumab (Arzerra®), such as abatacept (Orencia®) and anakinra (Kineret®) And belimumab (Benlysta®), CD4 biologics and other cytokine inhibitors or biological agents of T-cell or B-cell receptor or interleukin. Examples of suitable anti-TNF agents include anti-TNF Biologics such as Enbrel® (etanecerpt), Humira® (adalimumab), Remicade® (English) Leisy monoclonal antibody (infliximab)), Cimzia® (certolizumab (certolizumab)) and Simponi® (golimumab (golimumab)).

Examples of other suitable anti-inflammatory agents include 5-aminosalicylic acid and mesalamine preparations, sulfasalazine, hydroxychloroquine, thioindole (azathioprine, fluorenyl hydrazide), methylamine oxime, ring Phosphonamine, cyclosporin, calcineurin inhibitors (cyclosporine, pimecrolimus, tacrolimus), mycophenolic acid (CellCept®), mTOR inhibitor (temsirolimus, everolimus), JAK inhibitor (tofatinib), (Xeljan®), Syk inhibitor (fostatinib) (fostamatinib)), corticosteroids, especially low dose corticosteroids (such as prednisone® and budesonide) and anti-inflammatory biologics such as anti-IL6R mAb (Actemra® (taciximab), anti-IL6) Biologics, anti-IL1 (Kineret®, canakinumab (Ilaris®), rilonacept (Arcalyst®)) or IL12 or IL23 biologics (Ute Stelara®, anti-IL17 biologics (secukinumab), anti-CD22 (epratuzumab), anti-integrin agents (that Natalizumab (Tysabri®), vedolizumab (Entyvio®), anti-IFNa (sifalimumab), anti-CD20 mAbs (rituximab (Rituxan®) ) and olfaximab (Arzerra®) and other agents such as Orencia®, Kineret®, Ilaris®, and Ellalyst ®), Securginumab, Ipaduzumab, Sifalimumab and belimumab (Benlysta®), CD4 biologics and other cytokine inhibitors or T-cells or B-cells Biological agent for receptor or interleukin. Suitable anti-TNF agents include anti-TNF biologics such as Enbrel®, Humira®, Remicade®, Cimzia®, and Cimzia®. Simponi® (golimumab). The invention also provides a compound of the invention for use in therapy. In particular, the invention provides a compound described herein, or a pharmaceutically acceptable salt thereof, for use in therapy. More particularly, the present invention provides 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazoline 4-Amine or a pharmaceutically acceptable salt thereof for use in therapy.

In additional embodiments, the invention provides a compound or a compound of the invention for use in a disease or condition for the treatment of a RIP2 kinase vector. In particular, the invention provides a compound, or a pharmaceutically acceptable salt thereof, as described herein for use in a disease or condition for the treatment of a RIP2 kinase vector.

In a further embodiment the invention provides a compound described herein or a pharmaceutically acceptable salt thereof for use in the treatment of uveitis, interleukin-1 converting enzyme-related fever syndrome, dermatitis, acute lung injury, Type 2 diabetes, arthritis (especially rheumatoid arthritis), inflammatory bowel disease (such as ulcerative colitis and Crohn's disease), early onset inflammatory bowel disease, extraintestinal inflammatory bowel disease, in response Prevention of ischemic reperfusion (especially kidney), liver disease (nonalcoholic steatohepatitis, alcoholic steatohepatitis and autoimmune hepatitis) in solid organs of ischemia caused by cardiac surgery, organ transplantation, sepsis and other injuries , allergic diseases (such as asthma), transplant reactions (such as graft versus host disease), autoimmune diseases (such as systemic lupus erythematosus and multiple sclerosis) and granulomatous disorders (such as sarcoidosis, Blau syndrome, early onset) Sexual sarcoidosis, Wegener's granulomatosis or interstitial lung disease).

In a further embodiment the invention provides a compound as described herein, or a pharmaceutically acceptable salt thereof, for use in the treatment of uveitis. In a further embodiment the invention provides a compound described herein, or a pharmaceutically acceptable salt thereof, for use in the treatment of a syndrome associated with interleukin-1 convertase. In a further embodiment the invention provides a compound described herein, or a pharmaceutically acceptable salt thereof, for use in the treatment of Blau syndrome. In a further embodiment the invention provides a compound as described herein, or a pharmaceutically acceptable salt thereof, for use in the treatment of early onset sarcoidosis. In a further embodiment the invention provides a compound as described herein, or a pharmaceutically acceptable salt thereof, for use in the treatment of ulcerative colitis. In a further embodiment the invention provides a compound as described herein, or a pharmaceutically acceptable salt thereof, for use in the treatment of Crohn's disease. In a further embodiment the invention provides a compound as described herein, or a pharmaceutically acceptable salt thereof, for use in the treatment of an early onset inflammatory bowel disease. In a further embodiment the invention provides a compound as described herein, or a pharmaceutically acceptable salt thereof, for use in the treatment of parenteral inflammatory bowel disease. In a further embodiment the invention provides a compound as described herein, or a pharmaceutically acceptable salt thereof, for use in the treatment of Wegener's granulomatosis. In a further embodiment the invention provides a compound as described herein, or a pharmaceutically acceptable salt thereof, for use in the treatment of sarcoidosis.

The invention also provides the use of a compound of the invention in the manufacture of a medicament for the treatment of a RIP2 kinase vector, such as the various diseases and conditions described herein. In particular, the invention provides the use of a compound described herein, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disease or condition of a RIP2 kinase vector. More particularly, the present invention provides 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazoline Use of a 4-amine or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of a disease or condition of a RIP2 kinase vector.

Accordingly, the present invention provides the use of a compound described herein, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a human in need thereof for a disease or condition mediated by RIP2 kinase. Therapeutic "effective amount" is intended to mean the amount of the compound which, when administered to a patient in need of such treatment, is sufficient to provide a therapeutic effect as defined herein. Thus, for example, a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof is an amount of a pharmaceutical agent of the present invention which, when administered to a patient in need thereof, is sufficient to modulate and inhibit RIP2 kinase activity. The symptoms of the disease mediated by the activity are reduced, alleviated or blocked. Equivalent to the amount of a particular compound in this amount, depending on various factors, such as the specific compound (eg, potency (pIC ₅₀ ), utility (EC ₅₀ ), and biological half-life of a particular compound), disease symptoms and severity, need for this The identity of the patient being treated (eg, age, size, and weight) can vary, but can be determined routinely by those skilled in the art. Similarly, the therapeutic efficacy of the compound and the period of administration (time between doses and timing of administration, such as pre-meal/meal/post-meal) will depend on the identity of the mammal being treated, the particular compound and its properties (eg, drug) The kinetic properties), the disease or condition and its severity may vary from the particular composition and method used, but may be determined by those skilled in the art.

"Treatment" is intended to mean at least ameliorating a disease or condition in a patient. Therapeutic methods for alleviating a disease or condition include the use of a compound of the invention in any conventionally acceptable manner, such as a disease or condition for preventing, delaying, preventing, treating or curing a vector. Specific diseases or conditions that are particularly susceptible to treatment using the compounds of the invention are described herein.

The compounds of the invention may be administered in any suitable route of administration, including both systemic and topical administration. Systemic administration includes oral administration, parenteral administration, transdermal administration, rectal administration, and administration by inhalation. Parenteral administration refers to a route of administration other than the intestinal, transdermal or inhalation, and which is typically by injection or infusion. Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion. Inhalation refers to administration to the lungs of a patient, whether through the mouth or through the nasal passages. Topical administration includes application to the skin.

The compounds of the invention may be administered once or according to a given regimen wherein a plurality of doses are administered at different time intervals during a period of administration. For example, the dose can be administered one, two, three or four times a day. The dosage can be administered until the desired therapeutic effect is achieved or the desired therapeutic effect is maintained indefinitely. Suitable dosage therapies for the compounds of the invention will depend on the pharmacokinetic properties of the compound, such as absorbency, distribution and half-life, as determined by the skilled artisan. In addition, suitable dosage therapies, including the duration of administration of the therapy, are in accordance with the severity of the disease or condition, disease or condition to be treated, and the condition to be treated, in the compounds of the present invention. The age and physical condition, the medical history of the patient to be treated, the nature of the concurrent treatment, the desired therapeutic effect, and the like are determined by the skill and knowledge of the skilled practitioner. It will be further appreciated by those skilled in the art that this dosage therapy may require adjustment when it is responded to by individual patients or when individual patients need to change over time.

For use in therapy, the compounds of the invention are generally, but not necessarily, formulated into a pharmaceutical composition prior to administration to a patient. Accordingly, the invention also relates to pharmaceutical compositions comprising a compound of the invention and one or more pharmaceutically acceptable excipients.

In one embodiment, the invention provides 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazoline A pharmaceutical composition of a 4-amine or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients. In a further embodiment, the invention provides 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazoline A pharmaceutical composition of a 4-amine (as a free base) and one or more pharmaceutically acceptable excipients. In a further embodiment, 6-(T-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazole) is provided which comprises a crystalline form characterized by the PXRD pattern of Figure 1. A pharmaceutical composition of 3-yl)-7-ethoxyquinazolin-4-amine and one or more pharmaceutically acceptable excipients. The pharmaceutical compositions of the present invention can be prepared and packaged in a wide variety of forms, wherein an effective amount of a compound of the present invention can be extracted and then administered to a patient, for example, in powders, syrups, and injectable solutions. Alternatively, the pharmaceutical compositions of the present invention can be prepared and packaged in unit dosage form. For oral administration, for example, one or more lozenges or capsules may be administered. A dose of the pharmaceutical composition contains at least one therapeutically effective amount of a compound of the invention. When prepared in unit dosage form, the pharmaceutical compositions may contain from 1 mg to 1000 mg of a compound of the invention.

As provided herein, a unit dosage form (pharmaceutical composition) containing from 1 mg to 1000 mg of the compound of the present invention can be administered one, two, three or four times a day, preferably one, two or three times per day, and more preferably one per day or Two times to achieve the effect of a disease or condition that treats the RIP2 vector.

The pharmaceutical composition of the present invention typically contains a compound of the invention. However, in a specific embodiment, the pharmaceutical composition of the present invention contains more than one of the inventions. Compound. Furthermore, the pharmaceutical compositions of the present invention may further comprise one or more additional pharmaceutically active compounds as desired.

As used herein, "pharmaceutically acceptable excipient" means a substance, composition or vehicle that is involved in imparting a form or firmness to a composition. When mixed, the excipients must be compatible with the ingredients of the other pharmaceutical compositions such that they are administered to the patient to avoid interactions which substantially reduce the potency of the compounds of the invention and which may result in pharmaceutical compositions being pharmaceutical Unacceptable interaction forces. In addition, each excipient must of course be of such high purity that it is pharmaceutically acceptable.

The compounds of the invention and the pharmaceutically acceptable excipients will typically be formulated in a dosage form suitable for administration to the patient in the desired route of administration. The conventional dosage forms include such suitable (1) oral administration, such as tablets, capsules, caplets, tablets, buccal tablets, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets, sachets; (2) parenteral administration, such as sterile solutions, suspensions, dispersions for reconstitution; (3) transdermal administration, such as transdermal patches; (4) rectal administration, such as suppositories; (5) inhalation, For example, aerosols and solutions; and (6) topical administration, such as creams, ointments, lotions, solutions, pastes, syrups, foams, and gels.

Suitable pharmaceutically acceptable excipients will vary depending upon the particular dosage form chosen. In addition, suitable pharmaceutically acceptable excipients can be selected according to their particular function in the composition. For example, a particular pharmaceutically acceptable excipient can be selected for its ability to help produce a uniform dosage form. The particular pharmaceutically acceptable excipient can be selected for its ability to aid in the production of a stable dosage form. Particular pharmaceutically acceptable excipients may be selected for their ability to carry or carry a compound or a compound of the invention from one organ or part of the body to another organ or part of the body once administered to the patient. Specific pharmaceutically acceptable excipients may be selected for their ability to enhance patient compliance.

Suitable pharmaceutically acceptable excipients include the following types of excipients: diluents, fillers, binding agents, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents , cosolvents, suspending agents, emulsifiers, sweeteners, flavors, flavor masking agents, tune Colorants, anti-caking agents, humectants, chelating agents, plasticizers, tackifiers, antioxidants, preservatives, stabilizers, surfactants, and buffers. It will be appreciated by those skilled in the art that certain pharmaceutically acceptable excipients can be used as more than one function and can be used as additional in accordance with the amount of excipients present in the formulation and other ingredients present in the pharmaceutical composition. Features.

The skilled artisan has the knowledge and skill in the art to select the appropriate amount of suitable pharmaceutically acceptable excipient for use in the present invention. In addition, there are a number of sources describing pharmaceutically acceptable excipients and which can be used to select suitable pharmaceutically acceptable excipients for use by those skilled in the art. Examples include Remington's Pharmaceutical Sciences, The Handbook of Pharmaceutical Additives, and The Handbook of Pharmaceutical Excipients.

The pharmaceutical compositions of the present invention are prepared using techniques and methods known to those skilled in the art. Some of the methods commonly used in the art are described in Remington's Pharmaceutical Sciences.

In one aspect, the invention relates to solid oral dosage forms such as, but not limited to, lozenges or capsules comprising an effective amount of a compound of the invention and a diluent or filler. Suitable diluents and fillers include lactose, sucrose, glucose, mannitol, sorbitol, starch (eg, corn starch, potato starch, and pregelatinized starch), cellulose and its derivatives (eg, microcrystalline cellulose), sulfuric acid Calcium and calcium hydrogen phosphate. The oral solid dosage form can further comprise a binding agent. Suitable binders include starch (eg, corn starch, potato starch, and pregelatinized starch), gelatin, gum arabic, sodium alginate, alginic acid, tragacanth, Guanhua bean gum, polyvinylpyrrolidone (povidone), and fiber. And its derivatives (such as microcrystalline cellulose). The oral solid dosage form can further comprise a disintegrant. Suitable disintegrants include: interlinked polyvinylpyrrolidone, sodium starch glycolate, cross-linked carboxymethylcellulose, alginic acid, and sodium carboxymethylcellulose. The oral solid dosage form can further comprise a lubricant. Suitable lubricants include stearic acid, magnesium stearate, calcium stearate and talc.

Figure 1 shows 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazoline-4 in crystalline form. - X-ray powder diffraction (PXRD) pattern of amine (free base).

Figure 2 shows the compound 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazolin-4- The amine was pre-administered to the rat, followed by administration of L18-MDP, and the combined IL8 cytokine response in the obtained rat whole blood sample.

Instance

The following examples illustrate the invention. These examples are not intended to limit the scope of the invention, but rather provide a guide for those skilled in the art to make and use the compounds, compositions and methods of the invention. Various changes and modifications may be made without departing from the spirit and scope of the invention.

The invention also includes deuterated forms of the various compounds of the invention. Each of the available hydrogen atoms bonded to the carbon atom can be independently replaced by a deuterium atom. Those skilled in the art will know how to synthesize the deuterated form of the compounds of the invention.

Name-based intermediates and final compounds described herein of using from Advanced Chemistry Development, Inc., 110 Yonge Street, 14 th Floor, Toronto, Ontario, Canada, M5C 1T4 (http://www.acdlabs.com/) of The ACD/Name Pro V6.02 software naming program is generated from the ChemDraw, Struct=Name Pro 12.0 naming program from ChemSoftDraw Ultra, CambridgeSoft.100 CambridgePark Drive, Cambridge, MA 02140 USA (www.cambridgesoft.com).

In the following experimental instructions, the following abbreviations may be used:

Preparation 1 6-(T-butylsulfonyl)-7-fluoroquinazolin-4-ol

Step 1: 6-(Tertiary butylthio)-7-fluoroquinazolin-4-ol: 6-bromo-7-fluoroquinazolin-4-ol (69 g, 285 mmol), hydrazine (three A mixture of phenylphosphine)-palladium(0) (20 g, 17 mmol) and sodium carbonate (60 g, 570 mmol) was stirred in DMF (1 L) with nitrogen for 5 min. 2-Methylpropan-2-thiol (64 ml, 570 mmol) was added and the reaction mixture was heated at <RTI ID=0.0> The reaction was cooled and filtered through a glass filter paper, and then 1500 mL of stirred water was slowly poured. The resulting red precipitate was filtered and triturated with 200 mL EtOAc. The solid was filtered and subsequently washed with 110 mL hexanes, 150 mL of EtOAc:EtOAc:EtOAcield ) is a tan solid. LC/MS: M+H=253.2 ¹ H NMR (400 MHz, DMSO-d ₆ ) δ </ RTI></RTI> 12.23-12.72 (m, 1 H), 8.24 (d, J = 8.1 Hz, 1 H), 8.19 (s, 1 H), 7.58 (d, J = 9.6 Hz, 1 H), 1.28 (s, 9 H).

Step 2: 6-(T-butylsulfonyl)-7-fluoroquinazolin-4-ol: 6-(Tertiary butylthio)-7-fluoroquinazolin-4-ol (45 g , 124 mmol) and a suspension of oxone (191 g, 311 mmol) in ethyl acetate (1220 ml), methanol (1220 ml) and water (1220 ml) were stirred at 25 ° C for 4 h and then added 25 g (total 2.8 Eq) oxone. The reaction mixture was stirred with an overhead stirrer for 12 h. The reaction was filtered and the filtrate was slowly basified with saturated aqueous sodium bicarbonate and then basified to pH 7.5 with solid sodium hydrogen carbonate. The mixture was extracted with an additional 1.25 L of EtOAc then EtOAc EtOAc. The combined organic layers were washed with brine then dried over MgSO ₄ A small amount of impurities were removed by wet milling with 200 mL EtOAc. The desired 6-(t-butylsulfonyl)-7-fluoroquinazolin-4-ol (33.2 g, 94% yield) was filtered elute LC/MS: M+H=285.2 ¹ H NMR (400 MHz, DMSO-d ₆ ) δ </ RTI></RTI></RTI></RTI></RTI></RTI></RTI></RTI></RTI></RTI></RTI></RTI><RTIgt; (s, 1 H), 7.73 (d, J = 11.1 Hz, 1 H), 1.17- 1.40 (s, 9 H).

Preparation 2 6-(T-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-fluoroquinazolin-4-amine

To 6- (tert-butyl sulfonylurea yl) -7-fluoro-quinazolin-4-ol (4.14g, 14.56mmol) of acetonitrile (42.7 mL) was added POCl ₃ (2.036ml, 21.84mmol) and DIEA (3.81 ml, 21.84 mmol). The reaction was heated to 80 ° C for 16 h overnight. Additional POCl ₃ (500 uL) was added and the reaction was stirred at 80 ° C for 18 h. Complete conversion to chloride was observed via LCMS. 4,5-Dimethyl-1H-pyrazol-3-amine (1.942 g, 17.47 mmol) was added and the reaction was stirred at 80 ° C for 1 h. The precipitate was filtered, washed with acetonitrile and dried to give 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-fluoroquinazoline 4-Amine, hydrochloride (4.15 g, 9.93 mmol, 68.2% yield). ^{1 H NMR (400MHz, DMSO-} d 6) δ ppm 9.10-9.44 (m, 1 H) 8.88 (br.s., 1 H) 7.94 (d, J = 10.36Hz, 1 H) 2.23 (s, 3 H ) 1.82(s,3 H)1.24-1.45(m,9 H). MS(m\z)378(M+H)+.

Example 1 6-(T-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-(2-methoxyethoxy)quinazoline-4 -amine

6-(T-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-fluoroquinazolin-4-amine, hydrochloride (300 mg A mixture of 2-methoxyethanol (5.7 ml, 73 mmol) and KOtBu (410 mg, 3.6 mmol) was stirred at 90 ° C for 4 d. The reaction was concentrated to dryness, dried and purified by chromatography on silica gel loaded (ISCO-Rf, 0-25% methanol (via with 1% NH ₄ OH) / ethyl acetate to give 6- (tert-butyl sulfo acyl -N-(4,5-Dimethyl-1H-pyrazol-3-yl)-7-(2-methoxyethoxy)quinazolin-4-amine (230 mg, 0.531 mmol, 73.2%) yield) as a yellow ^{solid. 1 H NMR (400MHz, DMSO} -d6) δ ppm 12.19 (s, 1 H), 10.36 (s, 1 H), 8.99 (s, 1 H), 8.45 (s, 1 H) , 7.34 (s, 1 H), 4.26-4.42 (m, 2 H), 3.73 (t, J = 4.4 Hz, 2 H), 3.34 (s, 3 H), 2.18 (s, 3 H), 1.74 ( s, 3 H), 1.33 (s, 9 H). MS (m/z) 434.

Example 2 6-(T-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazolin-4-amine

Step 1: 6-(T-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-fluoroquinazolin-4-amine: at 6- Addition of POCl ₃ (2.70 ml, 29.0 mmol) and TEA (4.0 ml) to a suspension of (t-butylsulfonyl)-7-fluoroquinazolin-4-ol (5.50 g, 19.35 mmol) in acetonitrile (48 ml). , 29mmol). The reaction was stirred at 80 ° C until overnight. 4,5-Dimethyl-1H-pyrazol-3-amine (2.58 g, 23.2 mmol) was added to the solution and the mixture was stirred at &lt A solid began to precipitate. The reaction mixture was allowed to cool to room temperature. The solid was filtered and washed with cold acetonitrile. The solid was dried in vacuo to give 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-fluoroquinazolin-4-amine. Hydrochloride (4.91 g, 11.86 mmol, 61.3% yield). (M+H)+ 378.2.

Step 2: 6-(T-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazolin-4-amine: Sodium ethoxide (24 ml, 65.6 mmol, 21% in EtOH) and 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7- Fluoroquinazolin-4-amine, hydrochloride (4.80 g, 11.60 mmol) was mixed, and the suspension was heated at 80 ° C for 2 hr. The reaction mixture was allowed to cool to room temperature. EtOH was evaporated and the residue was dissolved in water (25 mL). The solution was neutralized to pH ~ 9 by the addition of 1 N HCl. A pale yellow solid precipitated. The solid was filtered, washed with water and dried in a vacuum oven overnight to give 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)- 7-Ethoxyquinazolin-4-amine (3.90 g, 9.67 mmol, 83% yield). (M+H) ⁺ 404.1; ¹ H NMR (DMSO-d ₆ , 400 MHz): δ=12.20 (s, 1 H), 10.36 (s, 1 H), 8.99 (s, 1 H), 8.46 (s, 1 H), 7.30 (s, 1 H), 4.13-4.34 (m, 2 H), 2.18 (s, 3 H), 1.74 (s, 3 H), 1.40 (t, J = 6.9 Hz, 3 H) , 1.33ppm(s, 9 H).

A sample of 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazolin-4-amine (120 g) ) suspended in EtOH (2000 ml) and then heated to 70 °C. An additional EtOH (2000 ml) was added and the resulting mixture was heated to reflux. Most of the solids are dissolved in the solvent. The hot suspension was filtered and poured into 12 L of cold water. The mixture was stirred for about 60 min and then allowed to stand overnight to bring the bath temperature to RT. The pale yellow precipitate was separated by filtration and dried in a vacuum oven to give 105.9 g (261 mmol, 88% yield) of crystal 6-(t-butylsulfonyl)-N-(4,5-dimethyl The group-1H-pyrazol-3-yl)-7-ethoxyquinazolin-4-amine is characterized by the PXRD mode of Figure 1, and the diffraction data of Table 1.

PXRD analysis was performed on a Rigaku Desktop X-ray diffractometer, Miniflex II, serial number DD02652 using a Scintillator NaI (TI) detector. The acquisition conditions include: Cu _Kα irradiation (λ=1.54059Å), generator voltage: 30kV, generator current: 15mA, starting angle: 3.0°2θ, ending angle: 40.0°2θ, step: 0.04°2θ, each step time : 0.5 seconds. Samples were prepared using zero background technology (positive aid).

Example 3 6-(T-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-propoxyquinazolin-4-amine

6-(T-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-fluoroquinazolin-4-amine (1.5 g, 3.97 mmol) A mixture of propan-1-ol (17.85 ml, 238 mmol) and KOtBu (2.230 g, 19.87 mmol) was heated at 90 ° C for 21 h. The reaction was poured into a solution of diethyl ether - became cloudy - no precipitate. The mixture was neutralized with citric acid and extracted with EtOAc (1×) and 2-MeTHF (1×). The combined organic layers were washed with brine, dried in Na ₂ SO ₄ and concentrated to dryness to give a crude product, which was purified via HPLC (10-50% ACN / water, 0.1% TFA). The product-containing fractions were placed in the partitioned between EtOAc and saturated sodium bicarbonate, washed with brine, dried in Na ₂ SO ₄ and concentrated to dryness. The resulting residue was triturated with EtOAc and filtered to give 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-propoxy The quinazolin-4-amine (280 mg, 0.671 mmol, 16.87% yield) was obtained as a white solid. ^{1 H NMR (400MHz, DMSO-} d 6) δ ppm 12.19 (s, 1 H) 10.36 (s, 1 H) 8.99 (s, 1 H) 8.45 (s, 1 H) 7.29 (s, 1 H) 4.17 ( t, J = 6.19 Hz, 2 H) 2.18 (s, 3 H) 1.76-1.84 (m, 2 H) 1.74 (s, 3 H) 1.26-1.37 (m, 9 H) 1.07 (t, J = 7.45 Hz) ,3 H). MS(m/z)418.3(M+H)+

Example 4 6-(T-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-((tetrahydrofuran-2-yl)methoxy)quinazoline 4-amine

To a solution of (tetrahydrofuran-2-yl)methanol (148 mg, 1.45 mmol) in EtOAc (EtOAc) The solution was stirred at room temperature for 5 min. Then 6-(t-butylsulfonyl)-7-chloro-N-(4,5-dimethyl-1Hpyrazol-3-yl)quinazolin-4-amine (30 mg, 0.076 mmol) The reaction mixture was stirred at 80 ° C until overnight. Most of the DMF is removed in a vacuum. The crude product was purified by a biotage column (0 to 16% MeOH/DCM) to afford 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazole-3- 7-((Tetrahydrofuran-2-yl)methoxy)quinazolin-4-amine (40 mg, 0.084 mmol, 35% yield). ¹ H NMR (DMSO-d6) δ: 12.19 (br.s., 1H), 10.36 (br.s., 1H), 8.99 (s, 1H), 8.45 (s, 1H), 7.34 (s, 1H) , 4.21 (m, 3H), 3.77-3.87 (m, 1H), 3.65-3.76 (m, 1H), 2.18 (s, 3H), 2.00 (m, 2H), 1.79-1.90 (m, 2H), 1.75 (s, 3H), 1.32 (s, 9H). MS (m/z) 460.

Pharmaceutical composition Example A

Preparation of tablets using conventional methods and blending as follows: amount of ingredients per ingot

Instance B

Capsules are prepared using conventional methods and formulated as follows:

Biological analysis:

A fluorescence polarization binding assay was performed to quantify the interaction of the novel test compound in the ATP-binding pocket of RIPK2 by competition with the ATP-competing ligand of the fluorescein. Full length FLAG His-tagged RIPK2 was purified from the baculovirus expression system and used at the final assay concentration of diploid KD. Fluorescent cursor-based ligand (5-({[2-({-3-({4-[(5-hydroxy-2-methylphenyl)amino)-2-pyrimidinyl)amino)phenyl Carbonyl}amino)ethyl]amino}carbonyl)-2-(6-hydroxy-3-oxirane-3H- -9-yl)benzoic acid, prepared as described in WO2011/120025, was used at a final assay concentration of 5 nM. Both the enzyme and the ligand were prepared in a solution of 50 mM HEPES pH 7.5, 150 mM NaCl, 10 mM MgCl ₂ , 1 mM DTT and 1 mM CHAPS. Test compounds were prepared in 100% DMSO and 100 nL were dispensed into individual wells of a porous disk. Next, 5 ul of RIPK2 was added to the test compound at twice the final assay concentration and incubated for 10 min at rt. After incubation, 5 ul of the fluorescein ligand solution was added to each reaction at twice the final assay concentration and incubated for at least 10 min at rt. Finally, the sample is placed on an instrument that measures fluorescence polarization. Test compound inhibition was expressed as percent inhibition (%) of the internal assay control.

In terms of concentration / dose-response experiments, normalized data were fit to the system measured pIC ₅₀ combined with the use of conventional techniques. The pIC ₅₀ for measuring the mean average of a minimum of two of the experiments.

Continuous test, to Example 1 (7.5) and Example 3 (8.1) mean pIC ₅₀ generates the proposed compound slight changes.

FLAG His labeled RIPK2 preparation:

The full-length human RIPK2 (receptor interaction serine-desaminase 2) cDNA line was purchased from Invitrogen (Carlsbad, California, USA, Clone ID: IOH 6368, RIPK2-pENTR 221). Gateway ^® LR was cloned for downstream of site-specific recombination RIPK2 to the N-terminal FLAG-6His contained in the target vector pDEST8-FLAG-His6 according to the method described by Invitrogen. Transfected into Spodoptera frugiperda (Sf9) insect cells using Cellfectin® (Invitrogen) according to the manufacturer's protocol.

Sf9 cell lines were grown in growth medium at 27 ° C, 80 rpm in a growth medium of Excell 420 (SAFC Biosciences, Lenexa, Kansas, US; Andover, Hampshire UK) until a sufficient amount of bioreactor was inoculated. The cells were placed in a 50 liter working bioreactor (Applikon, Foster City, California, US; Schiedam, Netherlands) at 27 ° C, 30% dissolved oxygen and a perturbation rate of 60-140 rpm until the desired amount was reached , a cell concentration of about 3.7 xe 6 cells/mL. Cells were infected with baculovirus at a multiplicity of infection (MOI) of 12.7. Training The lineage lasted for 43 hours. Infected cells were removed from the growth medium by centrifugation at 2500 g using Viafuge (Carr) continuous centrifugation at a flow rate of 80 liters per hour. The cell pellet was immediately frozen and subsequently used for purification.

Purification procedure I: 9.83 x 10 ⁵ insect cells were resuspended in 1.4 L of dissolution buffer (50 mM Tris (pH 8.0), 150 mM NaCl, 0.5 mM NaF, 0.1% Triton X-100, 1 mL / liter of protease inhibitor mixture group III (Available from EMD Group; CalBiochem/Merck Biosciences, Gibbstown, New Jersey, US; Damstadt, Germany) and treated on a Dunes cloud mill on ice. The suspension was then centrifuged at 47,900 g for 2 h at 4 °C. Clarification. The eluate was decanted from the insoluble mass and loaded onto a 55 mL FLAG-M2 affinity column (2.6 x 10.4 cm) at a linear flow rate of 16 cm/h, which has been buffered with 10 column volumes. Pre-equilibrated with liquid A (50 mM Tris (pH 8.0), 150 mM NaCl, 0.5 mM NaF, 1 mL/litre Protease Inhibitor Cocktail Set III). The column was then washed with 15 column volumes of buffer A and divided into 6 columns. Volume of Buffer B (buffer A + 150 μg / mL 3X FLAG peptide) was eluted at a linear flow rate of 57 cm / h. The fractions containing the relevant proteins were identified by SDS-PAGE, and the dialysis tube was folded using 10 kDa MWCO SnakeSkin to 5 L. Buffer A (without protease inhibitor cocktail) was dialyzed overnight to remove 3X FLAG peptide from the preparation. Health 11.3mg total protein, wherein the density measured by gel scanning, based RIPK2 40% purity is present, and to confirm the identity of the peptide mass fingerprint. Preparation major contaminating proteins was identified as a low molecular weight degradation fragments RIPK2.

Purification Procedure II: 100 g of cells (10 liters of fermentation) were frozen, thawed, and resuspended in 1 L of dissociation buffer (50 mM Tris HCL pH 7.5, 250 mM NaCl, 0.1 mM TCEP, 3 ml protease inhibitor cocktail) and homogenized at high pressure Dissociated at 10,000 psi (Avestin). The suspension was then centrifuged at 35,000 g for 45 minutes at 4 °C. The supernatant was centrifuged and incubated with 5 ml of anti-FLAG-M2 resin pre-equilibrated with buffer A (50 mM Tris HCL pH 7.5, 250 mM NaCl, 0.1 mM TCEP). After 1 hour of protein binding at 4 ° C, the resin was embedded in a 25 ml disposable column. Each column was washed with 25 ml of buffer A and dissolved in 10 ml (buffer A + 200 ug / ml Flag peptide). Concentrate the combined solution to 1 ml and apply to superdex The 200 (16/60) size excludes the column. The fraction containing the full length RIPK2 was collected based on the results of SDS-PAGE analysis. This purification yielded 36 mg/L of 80% pure RIPK2 protein and was identified by peptide quality fingerprinting.

Biological analysis:

A human tick hormone dipeptide (MDP)-stimulated human whole blood cytokine production assay was performed to assess the cellular potency and utility of the novel test compounds. Heparinized blood (160 μL) obtained from healthy human volunteers was dispensed into a well of a porous disk. The test compound was dissolved in 100% DMSO and diluted with D-PBS without calcium and magnesium to prepare a 10x working storage solution. Twenty microliters of the diluted test solution was added to each well and the porous disk was placed in a disk shaker (500 rpm) and incubated in a humidified incubator for 30 min (37 ° C, 5% CO ₂ ). A sterile, endotoxin-free 10% MDP working stock solution containing 1% DMSO was prepared. Twenty microliters of MDP stock solution was added to each well (final concentration = 100 ng/mL) to stimulate RIP2 kinase-dependent cytokine production. The final concentration of DMSO in all wells was 0.1% (v/v). The porous disk was incubated for an additional 6 hr (as noted above). An additional 100 μL of D-PBS (Dulbecco's Phosphate-buffered saline) per well was then added, the porous disk was centrifuged, and the supernatant was collected. The amount of TNFα in the supernatant was quantified using a commercially available immunoassay (Meso Scale Discovery). Test compound inhibition was expressed as percent inhibition (%) of the internal assay control. For concentration/dose response experiments, normalized data were fitted and pIC50 was determined using conventional techniques. The pIC ₅₀ for measuring the mean average of a minimum of two of the experiments.

Biological in vivo analysis - inhibition-induced inflammatory response

The utility of RIP2 inhibitors can also be assessed in vivo in rodents. Administration of L18-MDP intraperitoneally (ip) or intravenously (iv) in mice has been shown to induce an inflammatory response via the activation signal NOD2 delivery pathway (Rosenweig, HL, et al. 2008. Journal of Leukocyte Biology 84: 529 -536). Using conventional techniques, by measuring the amount of one or more cytokines (IL8, TNFα, IL6, and IL-1β) in serum and/or peritoneal washes or by measuring the flow into the peritoneal neutrophils (when L18- MDP was administered with ip to monitor the extent of inflammatory response in L18-MDP treated rats. In treated rats, inhibition of the L18-MDP-induced inflammatory response can be visualized by oral administration of the test compound, and then the amount of one or more cytokines in the serum (IL8, TNFα, IL6, and IL) is measured and compared using conventional techniques. -1β) to control treated animals.

For example, the compound of Example 2, 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl), at doses of 0, 0.04, 0.4, and 4 mg/kg -7-Ethoxyquinazolin-4-amine, pre-administered to rats, followed by administration of L18-MDP (50 μg/rat) for 0.25 h/min after pre-administration. In this study, the amount of IL8 cytokine in whole blood samples taken from rats was measured using antibody-based detection (Meso-Scale Discovery platform). The IL8 cytokine response was calculated as the mean response of each dose relative to the response observed in vehicle-treated rats, and is the mean ± standard deviation of the mean as shown in Figure 2 (n=8). Rat/group).

Biological in vivo analysis - rabbit heart wedge preparation

Female rabbits weighing 2.2-3 kg were anticoagulated with heparin and anesthetized with pentobarbital (50 mg/kg, i.v.). The thoracic cavity was opened via left thoracotomy and the heart was removed and placed in a cardiac arrest solution consisting of cold (4 °C) normal Tyrode's solution. A transmural wedge having a size of about 1.5 cm wide and 2-3 cm long was cut from the left ventricle.

The wedge tissue is inserted into the catheter via the left anterior descending artery or circumflex artery and perfused with a cardiac arrest solution. The preparation was then placed in a small tissue bath and perfused with arterial solution (T: 35.7 ± 0.1 ° C, perfusion pressure: 30-45 mm Hg). Let the ventricle wedge balance in the tissue fluid until electricity Stable, usually 1 hour. The preparation was incubated with a base cycle time (BCL) of 1000 to 2000 milliseconds, insulated with a bipolar silver electrode (except for the tip), and applied to the endocardial surface.

Use an extracellular silver/silver chloride electrode placed in a bench solution of the bath preparation, from the epicardium to the endocardial surface 1.0 to 1.5 cm, along the same vector as a transmembrane record (Epi: "+" Pole), recording the cross-wall electrocardiogram (ECG) of all experiments. On the ECG, the repolarization distribution (TDR) across the muscle layer is defined by the interval between the end of the T wave (T _pe ) and the peak. The QT interval is defined as the time from the start of the QRS to the last downhill point of the T wave of the wire. During the QRS, QT and Tp-e periods, 10 cycles were measured and averaged for each treatment. The data for each treatment of the entire fauna was averaged and compared to the average control value.

Isotonic contraction force generation (% ICF) measures 10 cycles and averages each treatment. The data for each treatment of the entire fauna was averaged and compared to the average control value.

Each test compound was prepared in 100% DMSO at a stock concentration of 30 mM. The compound was diluted to the highest concentration tested in Tyrode's buffer (containing mM: 129 NaCl, 4 KCl, 0.9 NaH ₂ PO ₄ , 20 NaHCO ₃ , 1.8 CaCl ₂ , 0.5 MgSO ₄ and 5.5 glucose, pH 7.4 when 95%) When O ₂ and 5% CO _{2 were} buffered, successive dilutions were subsequently prepared therefrom.

Each test compound was tested at 4 concentrations from 1-30 μM. After the wedge preparation was perfused with normal Tyrode's solution and stimulated with 1000 milliseconds of BCL for 1 hour, the stimulation frequency was reduced to a stable period of 5 minutes for BCL of 2000 milliseconds, after which baseline ECG and isotonic contraction (ICF) were recorded. The preparation was then returned to 1000 minutes of BCL and perfused with a Tyrode solution containing the test compound. For each test compound concentration, the wedge preparation was perfused with 1000 minutes of BCL for 20 minutes followed by a 2000 minute BCL perfusion for 5 minutes during which ECG and ICF were recorded. The compound of Example 2 (6-(T-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxy was evaluated in a rabbit heart wedge preparation. Quinazoline-4-amine). The four major readings from wedge preparations include QT prolongation, torsadegenicity (TdP scores derived from QT, Tp-e and early post-depolarization), and pulse conduction (QRS-related) And shrinkage, which is shown in Table 2.

A scoring system was used to assess the risk of the compound using the isolated rabbit left ventricular wedge preparation for relative TdP risk: QT interval score, _pe / QT ratio. The TdP score was generated by first converting the QT interval and the Tp-e/QT ratio to a % change from baseline. These values are assigned a TdP score based on the following system: <-5%=-1, -5% to 10%=0, 10% to 20%=1, 20% to 30%=2, >30% =3. The total score system range is -2 to 14 at BCL=2000ms.

QT = QT interval, Tp-e = discrete across the muscle layer, ICF = contractility.

Kinome Selectivity

Example 2 Compound (6-(T-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazolin-4-amine) The kinase group selectivity (as performed by Reaction Biology Corporation, One Great Valley Parkway, Malvern, PA, USA, 19355, http://www.reactionbiology.com) is characterized by in vitro characterization of the 337 member kinase group. Determination. It was verified that the compound of Example 2 at a concentration of 1 μM inhibited 1 of the 337 test kinases by >70% and inhibited 4 of the 337 test kinases by >50%. References: WO2011/120025, WO2011/120026, WO2011/123609, WO2011/140442, WO2012/021580, WO2012/122011, WO2013/025958.

Claims (15)

a compound having the formula: or Or its salt. a compound which is 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazolin-4-amine . a compound which is 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazolin-4-amine , or a pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazoline-4 An amine or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients. A pharmaceutical composition comprising 6-(t-butylsulfonyl)-N-(4,5-dimethyl-1H-pyrazol-3-yl)-7-ethoxyquinazoline-4 An amine and one or more pharmaceutically acceptable excipients. A compound of claim 2, or a pharmaceutically acceptable salt thereof, for use in therapy. A compound of the third aspect of the patent application is for use in therapy. A compound or a pharmaceutically acceptable salt thereof according to claim 2, for use in the treatment of a disease or condition of a RIP2 kinase vector. A compound according to claim 3, for use in a disease or condition for the treatment of a RIP2 kinase vector. A use of a compound of claim 2 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of a disease or condition of a RIP2 kinase vector. A compound according to claim 3, which is a disease or condition for the manufacture of a medicament for the treatment of a RIP2 kinase vector. The use of claim 10 or 11, wherein the disease or condition is selected from the group consisting of uveitis, interleukin-1 inducible enzyme-related fever syndrome, dermatitis, acute lung injury, type 2 diabetes, arthritis , rheumatoid arthritis, ulcerative colitis, Crohn's disease, early onset inflammatory bowel disease, parenteral inflammatory bowel disease, prevention of ischemic reperfusion injury in solid organ transplantation, nonalcoholic steatohepatitis, alcohol Sexual steatohepatitis, autoimmune hepatitis, asthma, graft versus host disease, systemic lupus erythematosus, multiple sclerosis, sarcoidosis, Blau syndrome, early onset sarcoidosis, Wegener's granulomatosis and interstitial lung disease . The compound of any one of claims 8 or 9 or a pharmaceutically acceptable salt thereof, wherein the disease or condition is selected from the group consisting of uveitis, interleukin-1 inducible enzyme-related fever syndrome, dermatitis, Acute lung injury, type 2 diabetes, arthritis, rheumatoid arthritis, ulcerative colitis, Crohn's disease, early onset inflammatory bowel disease, parenteral inflammatory bowel disease, prevention of ischemic in solid organ transplantation Reperfusion injury, nonalcoholic steatohepatitis, alcoholic steatohepatitis, autoimmune hepatitis, asthma, graft versus host disease, systemic lupus erythematosus, multiple sclerosis, sarcoidosis, Blau syndrome, early onset sarcoidosis , Wegener's granulomatosis and interstitial lung disease. The use of the invention of claim 10 or 11, wherein the disease or condition is selected from the group consisting of uveitis, a syndrome associated with interleukin-1 invertase, Blau syndrome, early onset sarcoidosis, ulcerative colitis, Crohn's disease, Wegener's granulomatosis and sarcoidosis. The compound of any one of claims 8 or 9 or a pharmaceutically acceptable salt thereof, wherein the disease or condition is selected from the group consisting of uveitis, interleukin-1 converting enzyme-related fever syndrome, Blau syndrome, Early onset sarcoidosis, ulcerative colitis, Crohn's disease, Wegener's granulomatosis, and sarcoidosis.